[1]陈颖慧 冯奕源 冯炜琦 吴逸卓 鲁亚南 于昱.血管发育异常的先天性心脏病患儿中Vav2基因突变的筛查和功能分析[J].心血管病学进展,2023,(8):757.[doi:10.16806/j.cnki.issn.1004-3934.2023.08.019]
　CHEN Yinghui,FENG Yiyuan,FENG Weiqi,et al.Identification and Functional Analysis of Vav2 Novel Variant in Congenital Heart Diseases with Vascular Malformation[J].Advances in Cardiovascular Diseases,2023,(8):757.[doi:10.16806/j.cnki.issn.1004-3934.2023.08.019]
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血管发育异常的先天性心脏病患儿中Vav2基因突变的筛查和功能分析()
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《心血管病学进展》[ISSN:51-1187/R/CN:1004-3934]

卷:
期数:
2023年8期
页码:
757
栏目:
论著
出版日期:
2023-08-25

文章信息/Info

Title:
Identification and Functional Analysis of Vav2 Novel Variant in Congenital Heart Diseases with Vascular Malformation
作者:
陈颖慧1 冯奕源 2 冯炜琦 1 吴逸卓 1 鲁亚南 3 于昱 1
(1.上海交通大学医学院附属新华医院心血管发育与再生医学研究所,上海 200092;2.上海交通大学医学院附属新华医院核医学科,上海 200092;3.上海交通大学医学院附属新华医院儿心脏中心,上海 200092)
Author(s):
CHEN Yinghui1; FENG Yiyuan2; FENG Weiqi1; WU Yizhuo1; LU Yanan3; YU Yu1
(1.Institute for Developmental and Regenerative Cardiovascular Medicine,Xinhua Hospital,School of Medicine,Shanghai Jiao Tong University,Shanghai 200092,China;2.Department of Nuclear Medicine,Xinhua Hospital,School of Medicine,Shanghai Jiao Tong University,Shanghai 200092,China;3.Department of Pediatric Cardiology,Xinhua Hospital,School of Medicine,Shanghai Jiao Tong University,Shanghai 200092,China)
关键词:
血管发育; 血管结构异常; 先天性心脏病; Vav2; 错义突变; 人脐静脉内皮细胞
Keywords:
Vascular development; Vascular malformation; Congenital heart disease; Vav2; Missense mutation; Human umbilical vein endothelial cell
DOI:
10.16806/j.cnki.issn.1004-3934.2023.08.019
摘要:
目的 通过全外显子组测序筛查血管结构异常类先天性心脏病（CHD）致病候选基因Vav2的新发突变c.701C＞T:p.P234L，探究Vav2及其新罕见突变对内皮细胞功能的影响，探索CHD发生的可能分子机制。方法 分析117例血管结构异常类CHD患儿及200例正常儿童的外周血全外显子组测序数据筛查Vav2突变。构建Vav2野生型及突变型表达质粒，转染人脐静脉内皮细胞（HUVEC），通过qRT-PCR和Western blotting检测Vav2和下游Rac1 mRNA和蛋白的表达水平；通过CCK-8、划痕实验和成管实验检测Vav2及突变对HUVEC增殖、迁移和成管能力的影响。结果 在血管结构异常类CHD患儿中发现的Vav2罕见突变c.701C＞T:p.P234L，在对照组中未出现且从未被报道。与野生型对比，P234L突变能降低Vav2蛋白表达，CCK-8、划痕实验和成管实验发现该突变影响Vav2促进HUVEC增殖、迁移和成管等血管生成相关的功能，但该突变不影响下游Rac1本底mRNA和蛋白表达。结论 Vav2的错义突变影响Vav2蛋白表达，进而损害Vav2在HUVEC中促进增殖、迁移和成管等血管生成相关的功能，可能是影响下游Rac1激活而非本底表达导致的。Vav2是血管发育过程的关键分子，参与调控血管结构异常类CHD发生，P234L突变导致Vav2功能受损，进一步提供了Vav2在血管结构异常类CHD中可能的致病分子机制。
Abstract:
Objective To identify novel variant c.701C＞T:p.P234L of pathogenic candidate gene Vav2 through Whole Exome Sequencing (WES),and investigate the effect of this variant on endothelial function to explore the pathogenesis of congenital heart diseases with vascular malformation (CHDs-VM). Methods The rare and novel Vav2 variant was identified in 117 CHDs-VM child patients and 200 healthy controls. Wild-type and variant Vav2 plasmids were constructed and transfected into human umbilical vein endothelial cells (HUVECs). mRNA and protein expression levels were detected by qRT-PCR and Western blot. CCK-8,wound healing assay and tube formation assay were used to analyze endothelial function alternations. Results The Vav2 variant c.701C＞T:p.P234L was identified in CHDs-VM patients but not observed in healthy controls. This variant down-regulated Vav2 protein expression and attenuated the promoting effect of Vav2 in proliferation,migration and tube formation of HUVECs. However,the mRNA and protein expressions of downstream Rac1 showed no difference between wild-type and variant transfected HUVECs. Conclusion The missense variant of Vav2 down-regulated Vav2 protein expressions,further impaired positive functions of Vav2 in proliferation,migration and tube formation of HUVECs probably through affecting downstream Rac1 activation rather than background expression. Deleterious variant induced Vav2 loss of function,which would uncover possible molecular mechanisms of Vav2 in CHDs-VM.

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更新日期/Last Update: 2023-09-21